JP2003066052A - Biological sample container - Google Patents

Abstract

(57) [Summary] [Problem] To provide a biological sample container capable of reducing the labor required when reacting and washing a large number of biological samples. SOLUTION: A biological sample container 1 in which a biological sample 3 is held in a plurality of recesses 4 is put into a tray 13 and a multi-dish 14 in which a necessary solution is put, and the space is formed by legs 9. The solution flowing in and out of the plurality of through-holes 7 provided in the bottom part 6 reacts and cleans the biological sample 3, so that even if the biological sample 3 is large, it is necessary to stick it to the slide glass. Thus, processes such as reaction and washing can be performed at once, and labor can be greatly reduced. In addition, the solution remaining in each recess 4 can be quickly discharged by lifting the biological sample container 1 from the solution surface without the need for a special suction operation or the like, so that the efficiency of the operation can be improved. .

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a biological sample container that reacts with a biological sample and is washed.

[0002]

2. Description of the Related Art Conventionally, a specific protein accumulation on a biological sample has been analyzed by an enzyme antibody method, or in situ.
In order to analyze the expression of a specific mRNA by the hybridization method, a biological sample is sliced and fixed on a slide glass.

Therefore, when the same chemical treatment is applied to all biological samples, a slide glass having the biological sample fixed is set up in a dedicated holder and the holders are dipped into a container such as a beaker containing the target chemical. Need to be processed.

[0004]

However, in order to fix a thin sliced sample of a living body on a slide glass, many treatments must be carried out even when using an embedding medium such as paraffin or by freezing. It takes a lot of time when the number of samples is large. Further, if one slide glass is used for one biological sample, a considerable space is required when the number of biological samples is large, and the analysis becomes complicated work.

Further, even when a plurality of different or various biological samples are fixed to one slide glass, it requires skill to fix them at equal intervals, and the subsequent chemical treatment becomes very troublesome.

[0006] When the slide glass is placed upright and placed in a container such as a beaker containing the target chemicals together with the holder, the biological sample may be peeled from the slide glass unless it is firmly fixed. .

The present invention has been made to solve the above problems, and provides a container for a biological sample capable of reducing the labor, etc., which occurs when a large number of biological samples are reacted or washed. Is intended.

[0008]

In order to achieve the above object, a biological sample container according to a main aspect of the present invention comprises:
At least a biological sample container for washing a biological sample, a plate, a plurality of recesses that are provided integrally with the plate and that have a plurality of through holes in each bottom and that hold the biological sample, and the recesses. And a leg portion that is provided integrally with each of the recesses on the outer side of the bottom portion and that supports each of the recesses.

In the present invention, while holding the biological sample in the plurality of recesses, the biological sample is reacted and washed with various solutions flowing in and out from the plurality of through holes provided in the bottom through the space formed by the legs. Therefore, a large number of biological samples can be treated at one time such as reaction and washing, and the labor can be greatly reduced. Moreover, since the solution remaining in each recess can be promptly discharged by lifting the biological sample container from the surface of the solution without any special suction work, the efficiency of the work can be improved.

Further, since the plate, the concave portion having a plurality of through holes and the leg portion are integrally formed, even if various solutions are used at various temperatures, the connecting portions of the respective portions do not separate.
In particular, if the bottoms of the recesses are separate, there is a risk of rupture at the connection and the biological sample may flow out from the recesses when the sample is small. Not that. Furthermore, the number of manufacturing steps is small, and the manufacturing cost is low.

According to one aspect of the present invention, the through hole has an inner diameter of 0.5 mm to 1.0 mm. As a result, even a small biological sample will not fall out of the recess through the through hole, and the inflow and outflow of the solution from the through hole will be smooth.

According to one aspect of the present invention, the inner diameter of the through hole inside the bottom portion is approximately 0.8 mm, and the inner diameter outside the bottom portion is approximately 1.0 mm. This makes it possible to minimize the contact of the solution with the solution in the through-hole when the solution is discharged, and it is possible to more smoothly discharge the solution while preventing the biological sample from flowing out from the recess.

According to one embodiment of the present invention, the inner diameter of the through hole inside the bottom portion is approximately 1.0 mm, and the inner diameter outside the bottom portion is approximately 0.8 mm. As a result, the inflow of the solution can be made smoother while preventing the outflow of the biological sample from the recess.

According to one aspect of the present invention, the height from the lower end of the leg portion to the outside of the bottom portion is approximately 1 mm. Thus, for example, when the solution is put in another container such as a saucer and the biological sample put in each recess is washed, the solution is put in each container while reducing the amount of the solution put in the other container. The biological sample can be sufficiently immersed in the solution. Moreover, since there is a sufficient space under the through hole of each recess, the solution can be smoothly discharged from the recess.

According to one aspect of the present invention, the recess is
It is characterized in that they are provided at equal intervals by the arrangement in the row direction and the column direction. Thus, for example, if 6 recesses are provided in the row direction, the solution can be rapidly dispensed by using a multiwell pipette for 12 wells, so that a large amount of sample can be processed.

According to one embodiment of the present invention, the plate, the recess and the leg are made of polypropylene. This improves the resistance to the solution used for washing the biological sample and the like.

According to one aspect of the present invention, a surfactant is mixed in the plate, the recess and the leg. As a result, the solution can more smoothly flow out and flow in from the through holes provided in the recesses, and the efficiency of various analysis operations can be improved.

According to one aspect of the present invention, the plate, the recess and the leg are made of ABS resin. As a result, while simplifying the manufacturing process, it is possible to make the solution flow out and in through the through holes provided in each recess more smoothly, and to improve the efficiency of various analysis work.

According to one aspect of the present invention, a convex portion is provided at an upper edge portion of the concave portion. This allows
Even when different solutions are used in the adjacent recesses, it is possible to prevent the solutions from being mixed.

[0020]

BEST MODE FOR CARRYING OUT THE INVENTION Embodiments of the present invention will be described below with reference to the drawings.

FIG. 1 is an overall perspective view showing a biological sample container according to an embodiment of the present invention, FIG. 2 is a plan view of the biological sample container shown in FIG. 1, and FIG. 3 is a sectional view taken along line AA of FIG. It is a figure.

As shown in these drawings, the biological sample container 1 has, for example, a long side of about 132 mm and a short side of about 86 m.
On the surface of the rectangular plate 2 of m, a plurality of bottomed cylindrical recesses 4 for holding the biological sample 3 are formed, for example, in the row direction.
24 pieces are provided in an array of 4 pieces at equal intervals in the column direction. As a result, the wells of the 24-well multiple well plate for cell culture, which are generally commercially available, can have the same spacing and the same number, so that a 12-well multipipettor can be used. As a result, the solution can be rapidly dispensed and a large amount of sample can be processed.

Further, a convex portion 5 having a height of approximately 0.5 mm is provided on the circumference which is the upper edge portion of each concave portion 4,
It is difficult for the solution used in the adjacent recesses 4 to mix. Furthermore, a through hole 7 is provided in the bottom portion 6 of each recess 4,
In addition, on the bottom outer side 8 of each recess 4, a leg 9 supporting each recess 4 is provided.
Is provided.

The plate 2, the recesses 4 and the legs 9 constituting the biological sample container 1 are made of polypropylene, for example, and are integrally manufactured by, for example, press molding. By being made of polypropylene in this way, it is possible to improve resistance to a solution in washing and the like.

The polypropylene is mixed with a surfactant such as a fatty acid ester. As a result, the contact angle on the surface of the through hole 7 provided in the recess 4 is made small, and the inflow and outflow of the solution through the through hole 7 are made smoother as described later.

Further, instead of polypropylene mixed with a surfactant such as a fatty acid ester, an ABS resin (acrylonitrile butadiene styrene resin) is used to form the plate 2, the recess 4 and the leg 9 which constitute the biological sample container 1. For example, it can be integrally manufactured by press molding.

As a result, while simplifying the manufacturing process,
The outflow and inflow of the solution from the through hole 7 provided in each recess 4 can be further smoothed, and various analysis work can be made efficient.

Here, FIG. 4 is an enlarged plan view of the above-mentioned concave portion 4, and FIG. 5 is a sectional view taken along line BB of FIG.

As shown in these figures, each recess 4 has a disk-shaped bottom 6 having an inner diameter of about 11 mm and a height of about 17 on the outer circumference.
A side wall portion 10 of mm is provided upright. Each through hole 7 has a conical shape in which the inner diameter of the bottom inner side 11 is approximately 0.8 mm and the inner diameter of the bottom outer side 8 is approximately 1.0 mm. This makes it possible to minimize the contact of the solution or the like with the solution or the like in the through-hole 7 when the solution or the like is discharged, and even if the biological sample 3 is extremely small, the solution or the like that has flowed into the recess 4 while holding the biological sample 3 therein. Can be discharged in a short time.

The legs 9 provided on the bottom outer side 8 of each bottom 6 have, for example, a height from the lower end 12 of the leg 9 to the bottom outer side 8 of 1 mm and a lateral width of 2 mm. It is provided. By providing the legs 9 on the bottoms 6 in this manner, the solution that has flowed into the recess 4 when the recess 4 is pulled up from the solution can be quickly discharged.

Next, an example of how to use the biological sample container 1 configured as described above will be described.

In s for a partial section of human gastric mucosa
In the in-hybridization, first, a part of human gastric mucosa is sampled, and it is fixed by reducing it to 5 mm square. After that, the fixed one is embedded in an embedding medium such as a suitable polymer assembling agent, and further sliced with a microslicer or the like to a thickness of about 50 μm to prepare a biological sample 3.

Next, an RNA probe is prepared. In this case, DNA of various gene groups expressed in human gastric mucosa
Using the fragment as a template, a non-radioactive labeled RNA probe prepared in vitro is used.

Further, a proteolytic enzyme treatment is carried out. Also in this case, various solutions are put in the tray 13, and the biological sample container 1 is put in the tray 13 so that the biological samples 3 respectively put in the respective recesses 4 of the biological sample container 1 are immersed in the respective solutions. Also in this case, the biological sample container 1 can process 24 biological samples 3 at a time.
Moreover, it is possible to promptly discharge the solution remaining in each recess 4 by lifting the biological sample container 1 from the solution surface without any special suction work.

Further, after the treatment with a proteolytic enzyme, acetylation is performed in order to prevent the background due to nonspecific adsorption of the probe to the biological sample 3. Also in this case, 24 biological samples 3 can be acetylated at one time by putting the biological sample 3 that has undergone the proteolytic enzyme treatment together with the biological sample container 1 into the tray 13 containing a predetermined solution. Moreover, it is possible to promptly discharge the solution remaining in each recess 4 by lifting the biological sample container 1 from the solution surface without any special suction work.

Next, washing is performed to wash out the solution used for acetylation. In this case as well, the biological sample container 1 is placed in the tray 13 containing the washing solution, moved up and down about 10 times, and then left standing in the solution for 10 minutes. This is repeated twice, but in this case as well, the biological sample container 1 can process 24 biological samples 3 at a time. Moreover, it is possible to promptly discharge the solution remaining in each recess 4 by lifting the biological sample container 1 from the solution surface without any special suction work.

Further, prehybridization is carried out to improve the penetration of the probe and suppress non-specific adsorption. That is, the biological sample container 1 is placed in a tray 13 containing 50% formamide / 5 × SSC, moved up and down about 10 times, and then left standing in the solution for 1 hour. Also in this case, the biological sample container 1 can process 24 biological samples 3 at a time. Moreover, it is possible to promptly discharge the solution remaining in each recess 4 by lifting the biological sample container 1 from the solution surface without any special suction work.

Next, an appropriate amount of RNA probe prepared for hybridization is added to the biological sample 3 contained in each recess 4. In this case, the multi-dish 14 shown in FIG. 7 may be used instead of the tray 13. In this case, not only one RNA probe can be added to each recess 4, but the same RNA probe can also be added. Also in this case, according to the biological sample container 1, it is not necessary to set a special chopstick on a square Petri dish like a slide glass, and arrange the slide glasses on the square chopsticks. Sample 3 will be processed.

After that, the plate is incubated for 16 hours and washed to remove the probe that has not hybridized.
That is, the multi-dish 14 is attached to the biological sample container 1
Then, the biological sample container 1 is placed in a tray 13 containing an appropriate amount of 50% formamide / 2 × SSC. At this time, dip for 20 minutes while keeping the temperature at the specified temperature. This process is repeated 3 times. In this case, the biological sample container 1
According to this, 24 biological samples 3 can be processed at one time. Moreover, it is possible to promptly discharge the solution remaining in each recess 4 by lifting the biological sample container 1 from the solution surface without any special suction work.

Furthermore, in order to lower the background due to non-specific binding, RNase treatment is performed. That is, the biological sample container 1 containing the washed biological sample 3 is placed in a tray 13 containing an appropriate amount of a predetermined solution, and formamide is washed away. In this case, according to the biological sample container 1,
This means that 24 biological samples 3 can be processed at one time. Moreover, it is possible to promptly discharge the solution remaining in each recess 4 by lifting the biological sample container 1 from the solution surface without any special suction work.

In order to remove the decomposition products, the biological sample container 1 containing the RNase-treated biological sample 3 is placed in the tray 13 containing an appropriate amount of a predetermined solution. This process is repeated 3 times. In this case, the biological sample container 1 can process 24 biological samples 3 at a time. Moreover, it is possible to promptly discharge the solution remaining in each recess 4 by lifting the biological sample container 1 from the solution surface without any special suction work.

Next, 100 mmol / liter Tris-
HCl (pH 7.5 at 20 ° C) and 150 mmol
The biological sample container 1 is placed in a tray 13 in which an appropriate amount of Buffer 1 made of NaCl / liter NaCl is put, the biological sample 3 is dipped therein, and the whole biological sample container 1 is moved up and down 10 times and then left still for 1 minute. In this case, the biological sample container 1 can process 24 biological samples 3 at a time. Moreover, it is possible to promptly discharge the solution remaining in each recess 4 by lifting the biological sample container 1 from the solution surface without any special suction work.

After this, 1% (W / V) blocking
The same process is performed with the tray 13 containing the Buffer 2 made by the reagent. Also in this case, according to the biological sample container 1, 24 biological samples 3 at a time
Can be processed. Moreover, it is possible to promptly discharge the solution remaining in each recess 4 by lifting the biological sample container 1 from the solution surface without any special suction work. Further, the processing by Buffer1 is repeated.

Next, an anti-DIG-AP labeled antibody is added to each biological sample 3 and allowed to stand at room temperature for 1 hour. Then Buff
Container 1 for biological sample is placed in tray 13 containing an appropriate amount of er1
Repeat the washing process to wash away excess antibody etc. twice, and further 100 mmol / liter Tris-H
Cl (pH 9.5 at 20 ° C.), 100 mmol / liter NaCl and 50 mmol / liter MgCl ₂
The same process is performed using Buffer 3 prepared by.

Finally, the NBT / BCIP color reaction solution is added to each biological sample 3 and allowed to stand in a dark room for 12 to 24 hours. In this case, according to the biological sample container 1, it is possible to surely perform processing without taking up much space as compared with the slide glass. When this reaction becomes clear with the lowest background and strong signal, 10 mmol / liter Tris-
The biological sample container 1 is placed in a tray 13 in which an appropriate amount of Buffer 4 made of HCl (pH 8.0 at 20 ° C.) and 1 mmol / liter EDTA is placed, and the reaction is stopped. Also in this case, according to the biological sample container 1,
This means that 24 biological samples 3 can be processed at one time. Moreover, it is possible to promptly discharge the solution remaining in each recess 4 by lifting the biological sample container 1 from the solution surface without any special suction work.

After this, a signal is usually present in the cytoplasm, and the nucleus appears to be missing. The identification of signals found in the nucleus and above the tissue is always done by comparison with the image of sense RNA hybridization to adjacent sections.

As described above, a partial section of human gastric mucosa
Although the use example of in situ hybridization has been described, the biological sample container 1 can be used for various methods, and as a biological sample 3, a small experimental animal, an entire individual such as an embryo, and a thick slice are sequentially analyzed for solution analysis. Anything can be used as long as it is soaked.

Therefore, in situ Hybrid
It can be used not only for zation, but also for antibody reaction, staining reaction, and various drug treatment experiments and drug screening on small individuals and embryos that survive in a solution system.

As described above, according to this embodiment, by placing the biological sample container 1 in which the biological samples 3 are held in the plurality of recesses 4 in the tray 13 and the multi-dish 14 containing the necessary solution, Since the solution that flows in and out from the plurality of through holes 7 provided in the bottom portion 6 through the space created by the portion 9 reacts, cleans, etc. the biological sample 3, there are cases where there are many biological samples 3. Even if it is not necessary to attach it to the slide glass, the reaction, washing and the like can be performed at one time, and the labor can be greatly reduced. In addition, since the biological sample container 1 can be lifted from the surface of the solution without any special suction work, the solution remaining in each recess 4 can be promptly discharged, so that the work efficiency can be improved. .

Further, since the plate 2, the concave portion 4 having the plurality of through holes 7 and the leg portion 9 are integrally formed, the connecting portions of the respective portions do not separate even if various solutions are used at various temperatures. In particular, if the bottom portion 6 and the side wall portion 10 of the concave portion 4 are separate, there is a risk that the joint portion may be split and the biological sample 3 may flow out from the concave portion 4 when the biological sample 3 is small. The biological sample container 1 does not have such a fear. Furthermore, the number of manufacturing steps is small, and the manufacturing cost is low.

Further, when processing slide glasses one by one, a large space is required if the number of biological samples 3 is enormous. However, the biological sample container 1 enables extremely compact processing and high work efficiency. Efficiency can be improved.

Further, since the height from the lower end 12 of the leg 9 of the biological sample container 1 to the bottom outer side 8 is set to about 1 mm, for example, when cleaning with the tray 13, the tray 13 is used.
It is possible to allow the biological sample 3 placed in each recess 4 to be sufficiently immersed in the solution while reducing the amount of the solution placed in the solution.

The present invention is not limited to the above-mentioned embodiments, and can be carried out by appropriately modifying it within the scope of the technical idea of the present invention.

For example, in the above-described embodiment, the inner diameter of the inner bottom portion 11 of the through hole 7 is 0.8 mm and the inner diameter of the outer bottom portion 8 is 1.0 mm.
Has an inside diameter of 1.0 mm and the outside 8 of the bottom has an inside diameter of 0.8 m
It may be m. Thereby, the inflow of the solution can be made smoother while preventing the outflow of the biological sample 3 from the recess 4.

Further, in the above-mentioned embodiment, the inner diameter of the bottom inner side 11 of the through hole 7 is 0.8 mm and the inner diameter of the bottom outer side 8 is 1.0 mm, but the inner diameter of the through hole 7 is 0.5 mm.
If the inside diameter of the through hole 7 is 0.5 mm and the inside diameter 11 of the bottom inside 11 is 0.5 mm, the inside diameter of the bottom outside 8 is 0.1 mm.
It may be 7 mm. Furthermore, the inside 11 of the bottom of the through hole 7
Has an inside diameter of 0.7 mm and the outside 8 of the bottom has an inside diameter of 0.5 m
It may be m.

Thereby, the smaller biological sample 3
Since the outflow from the through hole 7 can be prevented, the utilization range of the biological sample container is expanded.

Further, in the above-mentioned embodiment, the inner diameter of the inner bottom portion 11 of the through hole 7 is 0.8 mm and the inner diameter of the outer bottom portion 8 is 1.0 mm, but the inner diameter of the through hole 7 is 0.5 mm.
The cross-sectional shape of the through hole 7 may be cylindrical or the like as long as it is within the range of mm to 1.0 mm. As a result, even a small biological sample 3 will not fall out of the recess 4 from the through hole 7 and the solution will flow smoothly in and out of the through hole 7, and the work such as cleaning can be speeded up.

Further, in the above-mentioned embodiment, the concave portion 4 is composed of the disk-shaped bottom portion 6 and the cylindrical side wall portion 10, but the present invention is not limited to this, and for example, a polygonal plate-shaped bottom portion. 6 and a side wall portion 10 provided upright on the outer periphery thereof.

Further, in the above-described embodiment, 24 recesses 4 are provided in the row direction by 6 and the column direction by 4 at an equal interval. However, the present invention is not limited to this, and for example,
48 recesses 4 arranged in the row direction and 6 in the column direction at regular intervals, and 4 recesses 4 in the row direction and 12 in the column direction at 12 equal intervals What is provided may be used. This allows work with a higher degree of freedom.

Further, in the above-described embodiment, the leg portions 9 provided on the bottom outer side 8 of each bottom portion 6 are provided at two places, but the number is not limited to two places, and four places are also provided.
It may be in one place. With this, for example, in the case of one place, while ensuring a sufficient space under each bottom portion 6, the inflow and outflow of the solution through the through hole 7 are made smoother.

[0061]

As described above, according to the present invention,
It is possible to reduce the labor and the like generated when reacting, washing, etc. a large number of biological samples.

[Brief description of drawings]

FIG. 1 is an overall perspective view showing a biological sample container according to an embodiment of the present invention.

FIG. 2 is a plan view showing a biological sample container according to an embodiment of the present invention.

FIG. 3 is a cross-sectional view taken along the line AA showing the biological sample container according to the embodiment of the present invention.

FIG. 4 is an enlarged plan view showing a recess according to the embodiment of the present invention.

FIG. 5 is a BB cross-sectional view showing a recess according to the embodiment of the present invention.

FIG. 6 is a perspective view showing a tray according to the embodiment of the present invention.

FIG. 7 is a perspective view showing a multi-dish according to an embodiment of the present invention.

[Explanation of symbols]

1 Container for biological samples 2 plates 3 biological samples 4 recess 5 convex 6 bottom 7 through holes 8 bottom outside 9 legs 10 Side wall 11 Inside bottom 12 Lower end 13 trays 14 multi-dish

   ─────────────────────────────────────────────────── ─── Continued front page    F term (reference) 2G045 AA24 BB14 CB01 DA12 DA13                       DA14 DA36 FB02 FB03 FB07                       HA06 HA14 HA16                 2G052 AA28 AD26 DA06 EB12 FC06                       JA16                 2G058 AA09 BB15 CC02 FB02 GA01                 4B029 AA09 CC01 GB05 HA02 HA05

Claims (10)

[Claims] 1. A biological sample container for washing at least a biological sample, wherein: a plate; and a plurality of recesses that are integrally provided with the plate and that have a plurality of through holes in each bottom and that hold the biological sample. And a leg portion that is provided integrally with each of the recesses on the outer side of the bottom of each of the recesses and supports each of the recesses. 2. The biological sample container according to claim 1, wherein the through hole has an inner diameter of 0.5 mm to 1.0 mm. 3. The biological sample container according to claim 1, wherein the inner diameter of the through hole inside the bottom portion is approximately 0.8 mm, and the inner diameter outside the bottom portion is approximately 1.0 mm. A container for a biological sample, characterized by: 4. The biological sample container according to claim 1 or 2, wherein the inner diameter of the inside of the bottom of the through hole is approximately 1.0 mm and the inner diameter of the outer side of the bottom is approximately 0.8 mm. A container for a biological sample, characterized by: 5. Any one of claims 1 to 4
The biological sample container according to item 1, wherein the height from the lower end of the leg to the outside of the bottom is approximately 1
A container for a biological sample, which has a size of mm. 6. Any one of claims 1 to 5
Item 4. The biological sample container according to item 4, wherein the recesses are provided at equal intervals in an array in a row direction and a column direction. 7. Any one of claims 1 to 6
Item 5. The biological sample container according to item 1, wherein the plate, the recess, and the leg are made of polypropylene. 8. Any one of claims 1 to 7
Item 5. The biological sample container according to item 4, wherein a surfactant is mixed in the plate, the recess, and the leg. 9. Any one of claims 1 to 6
Item 10. The biological sample container according to item 4, wherein the plate, the recess, and the leg are made of ABS resin. 10. The biological sample container according to claim 1, wherein a convex portion is provided on an upper edge portion of the concave portion.